LOCUS       V001361                 9637 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V001361
VERSION     V001361
KEYWORDS    pLenti CMV Puro LUC (w168-1)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9637)
  AUTHORS   Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO,
            Campisi J, Yaswen P, Cooper PK, Kaufman PD.
  TITLE     A versatile viral system for expression and depletion of proteins in
            mammalian cells.
  JOURNAL   PLoS One. 2009 Aug 6;4(8):e6529.
   PUBMED   19657394
REFERENCE   2  (bases 1 to 9637)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9637)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2009-08-6"; volume: "4(8)"; pages: "e6529"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     505..529
                     /label="LNCX"
                     /note="Human CMV promoter, forward primer"
     protein_bind    613..637
                     /label="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             643..2292
                     /label="luciferase"
                     /note="firefly luciferase"
     protein_bind    complement(2302..2326)
                     /label="attB2"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2357..2945
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     promoter        2959..3458
                     /label="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             3479..4075
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             4216..4449
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4521..4655
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      4682..4817
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(4838..4856)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4866..4882)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      5024..5479
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5505..5609
                     /label="AmpR promoter"
     CDS             5610..6467
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      6641..7229
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     7383..7400
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     protein_bind    7517..7538
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7553..7583
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    7591..7607
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7615..7631
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        7652..7670
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7698..7924
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7925..8105
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8152..8277
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    8770..9003
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             9188..9232
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             9381..9422
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    9499..9615
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgactc tagtccagtg tggtggaatt
      601 ctgcagatat caacaagttt gtacaaaaaa gcaggctcca ccatggaaga cgccaaaaac
      661 ataaagaaag gcccggcgcc attctatccg ctggaagatg gaaccgctgg agagcaactg
      721 cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac agatgcacat
      781 atcgaggtgg acatcactta cgctgagtac ttcgaaatgt ccgttcggtt ggcagaagct
      841 atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga aaactctctt
      901 caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc gcccgcgaac
      961 gacatttata atgaacgtga attgctcaac agtatgggca tttcgcagcc taccgtggtg
     1021 ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct cccaatcatc
     1081 caaaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc gatgtacacg
     1141 ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtgcc agagtccttc
     1201 gatagggaca agacaattgc actgatcatg aactcctctg gatctactgg tctgcctaaa
     1261 ggtgtcgctc tgcctcatag aactgcctgc gtgagattct cgcatgccag agatcctatt
     1321 tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt ccatcacggt
     1381 tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt cttaatgtat
     1441 agatttgaag aagagctgtt tctgaggagc cttcaggatt acaagattca aagtgcgctg
     1501 ctggtgccaa ccctattctc cttcttcgcc aaaagcactc tgattgacaa atacgattta
     1561 tctaatttac acgaaattgc ttctggtggc gctcccctct ctaaggaagt cggggaagcg
     1621 gttgccaaga ggttccatct gccaggtatc aggcaaggat atgggctcac tgagactaca
     1681 tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg taaagttgtt
     1741 ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg cgttaatcaa
     1801 agaggcgaac tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa caatccggaa
     1861 gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat agcttactgg
     1921 gacgaagacg aacacttctt catcgttgac cgcctgaagt ctctgattaa gtacaaaggc
     1981 tatcaggtgg ctcccgctga attggaatcc atcttgctcc aacaccccaa catcttcgac
     2041 gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt
     2101 ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta
     2161 acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt
     2221 accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa gaagggcgga
     2281 aagatcgccg tgtaattcta gacccagctt tcttgtacaa agtggttgat atccagcaca
     2341 gtggcggccg ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2401 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2461 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2521 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2581 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2641 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2701 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     2761 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     2821 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     2881 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     2941 cccgcctgga attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
     3001 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
     3061 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
     3121 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
     3181 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat
     3241 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg
     3301 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg
     3361 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg
     3421 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat
     3481 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     3541 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     3601 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     3661 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     3721 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     3781 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     3841 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     3901 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     3961 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     4021 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg
     4081 cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg catctcgagg
     4141 gcccggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     4201 aagaaaaggg gggactggaa gggctagctc actcccaacg aagacaagat ctgctttttg
     4261 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     4321 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     4381 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     4441 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     4501 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
     4561 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4621 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
     4681 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4741 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4801 aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg
     4861 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     4921 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     4981 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     5041 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5101 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5161 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5221 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5281 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5341 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     5401 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5461 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5521 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5581 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5641 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5701 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5761 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5821 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5881 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5941 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6001 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6061 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6121 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6181 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6241 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6301 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6361 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6421 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6481 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6541 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6601 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6661 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6721 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6781 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6841 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6901 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6961 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7021 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7081 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7141 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7201 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7261 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7321 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     7381 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     7441 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     7501 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     7561 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     7621 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
     7681 tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac
     7741 gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga
     7801 agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg
     7861 acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat
     7921 aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     7981 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     8041 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     8101 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
     8161 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
     8221 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
     8281 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
     8341 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
     8401 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
     8461 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
     8521 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
     8581 gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg
     8641 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     8701 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     8761 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     8821 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
     8881 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     8941 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     9001 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     9061 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     9121 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     9181 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     9241 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     9301 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     9361 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
     9421 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
     9481 tcgacggtat cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     9541 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     9601 aaaattcaaa attttatcga taagcttggg agttccg
//
